Cloning of the Maltose Phosphorylase Gene fromBacillussp. Strain RK-1 and Efficient Production of the Cloned Gene and the Trehalose Phosphorylase Gene from…
| Autor: | Tsuneya Yatake, Tetsuji Tomita, Nozomu Yasutake, Yasushi Inoue, Shinsuke Miyoshi, Yoshie Oshima, Yoshie Yamamoto |
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| Rok vydání: | 2002 |
| Předmět: |
Bacillaceae
biology Bacillus amyloliquefaciens Organic Chemistry General Medicine Maltose Bacillus subtilis Molecular cloning biology.organism_classification Applied Microbiology and Biotechnology Biochemistry Molecular biology Trehalose Bacillales Maltose phosphorylase Analytical Chemistry chemistry.chemical_compound chemistry Molecular Biology Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 66:2594-2599 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.66.2594 |
| Popis: | The maltose phosphorylase (MPase) gene of Bacillus sp. strain RK-1 was cloned by PCR with oligonucleotide primers designed on the basis of a partial N-terminal amino acid sequence of the purified enzyme. The MPase gene consisted of 2,655 bp encoding a theoretical protein with a M r of 88,460, and had no secretion signal sequence, although most of the MPase activity was detected in the culture supernatant of RK-1. This cloned MPase gene and the trehalose phosphorylase (TPase) gene from Bacillus stearothermophilus SK-1 were efficiently expressed intracellularly under the control of the Bacillus amyloliquefaciens α-amylase promoter in Bacillus subtilis. The production yields were estimated to be more than 2 g of enzyme per liter of medium, about 250 times the production of the original strains, in a simple shake flask. About 60% of maltose was converted into trehalose by the simultaneous action of both enzymes produced in B. subtilis. |
| Databáze: | OpenAIRE |
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